Peptide-Protein Interactions Suggest That Acetylation of Lysines 381 and 382 of p53 Is Important for Positive Coactivator 4-p53 Interaction
Peptide-Protein Interactions Suggest That Acetylation of Lysines 381 and 382 of p53 Is Important for Positive Coactivator 4-p53 Interaction
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DOI：
10.1074/jbc.m110.205328
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发表时间：
2011-07-15
影响因子：
4.8
通讯作者：
Roy, Siddhartha
中科院分区：
文献类型：
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作者：
Debnath, Subrata;Chatterjee, Snehajyoti;Roy, Siddhartha
人转录正辅激活因子4（PC 4）激活几个p53依赖性基因。已经证明这是与p53直接相互作用的结果。先前，我们已经得出结论，PC 4主要通过其DNA结合C-末端的一半与p53的C-末端负调控结构域相互作用。用肽片段进行的NMR化学位移扰动研究表明，p53的氨基酸380-386对于与PC 4的相互作用至关重要。荧光各向异性和沉降速度的研究证实了这一点。一种由p53-（380-386）序列组成的肽，当与细胞穿透标签和核定位信号连接时，定位于细胞核并抑制荧光素酶基因从携带在p53依赖性启动子下的Luc基因的转染质粒表达。赖氨酸382/381的乙酰化使该肽与PC 4的结合增强了约一个数量级。核磁共振和突变研究表明，PC 4的丝氨酸73是识别p53的重要残基。分子间核Overhauser效应将天冬氨酸76置于赖氨酸381附近，表明PC 4残基73-76周围的区域对p53识别很重要。p53的380-386区域与PC 4的73-76残基周围区域相互作用，p53的赖氨酸382/381的乙酰化可能在调节p53-PC 4相互作用以及PC 4介导的p53靶基因激活中起重要作用。
The human transcriptional positive coactivator 4 (PC4) activates several p53-dependent genes. It has been demonstrated that this is a consequence of direct interaction with p53. Previously, we have concluded that PC4 interacts mainly with the C-terminal negative regulatory domain of p53 through its DNA binding C-terminal half. NMR chemical shift perturbation studies with peptide fragments indicated that amino acids 380-386 of p53 are crucial for interaction with PC4. This was verified by fluorescence anisotropy and sedimentation velocity studies. A peptide consisting of p53-(380-386) sequence, when attached to a cell penetration tag and nuclear localization signal, localizes to the nucleus and inhibits luciferase gene expression from a transfected plasmid carrying a Luc gene under a p53-dependent promoter. Acetylation of lysine 382/381 enhanced the binding of this peptide to PC4 by about an order of magnitude. NMR and mutagenesis studies indicated that serine 73 of PC4 is an important residue for recognition of p53. Intermolecular nuclear Overhauser effect placed aspartate 76 in the vicinity of lysine 381, indicating that the region around residues 73-76 of PC4 is important for p53 recognition. We conclude that the 380-386 region of p53 interacts with the region around residues 73-76 of PC4, and acetylation of lysine 382/381 of p53 may play an important role in modulating p53-PC4 interaction and as a consequence PC4 mediated activation of p53 target genes.